Preparation of orodispersible tablets of bosentan using xylitol and menthol as dissolution enhancers.

Bosentan is a drug used to treat pulmonary hypertension via dual endothelial receptor antagonism. Bosentan has a restricted oral bioavailability, a problem that's mostly due to poor solubility and hepatic metabolism. It is extensively used for the elderly and children who require a friendly dosage form like orodispersible tablets. So, the goal of this research work was to hasten the dissolution rate of bosentan to produce an orodispersible tablet with immediate drug release. Bosentan was exposed to ethanol-assisted kneading with a rise of xylitol or menthol concentrations (1:1 and 1:2 molar ratio of bosentan with excipient). In addition to observing the dissolution behavior, the resulting dry products were investigated using Fourier transform infrared spectroscopy (FTIR), differential thermal analysis (DTA), and X-ray diffraction (XRD). The FTIR reflected possible hydrogen bonding with xylitol and menthol. DSC studies reflected a reduction in the enthalpy and Tm. These results with XRD data reflected partial co-amorphization in the case of xylitol and eutaxia in the case of menthol. These modifications were related to an accelerated dissolving rate. The developed systems were fabricated as orodispersible tablets which exhibited immediate release of bosentan. Thus, the current study offered simple co-processing for the preparation of orodispersible bosentan tablets.

Developing of an eco-friendly liquid-liquid microextraction method by using menthol-based hydrophobic deep eutectic solvent for determination of basic fuchsin dye: assessment of the greenness profile.

Herein, we aimed to develop a new environmentally friendly liquid-liquid microextraction (LLME) method based on hydrophobic deep eutectic solvent (hDES) synthesized using biodegradable dl-menthol and decanoic acid for the spectrophotometric determination of toxic basic fuchsin dye in environmental water samples. The parameters affecting the extraction efficiency such as pH, mole ratio, and volume of hDES (1:2) and type and volume of organic solvent, sample volume, times of vortex, ultrasonic bath and centrifuge, ionic strength, and matrix effect were investigated and optimized. Under optimal conditions, the calibration curve showed linearity in the range of 7.4-167 µg L-1 with a coefficient of determination of 0.9994. The limit of detection, intra-day and inter-day precision, and recovery values were 2.25 µg L-1, 2.46% and 4.45%, and 105 ± 3%, respectively. The preconcentration and enrichment factors were found to be 30 and 61.5, respectively. The proposed hDES-LLME methodology was successfully applied to the environmental water samples to detect toxic BF dye (95-105%). Finally, the ecological impact of the suggested method was evaluated using the analytical eco-scale (PPS:88), complementary green analytical procedure indexe (ComplexGAPI), and the Analytical GREEnness tool (0.63). The assessment results showed that the presented analytical method can be regarded as a green LLME approach for the determination of the BF in water.

Enhancing the Enantioselectivity and Catalytic Efficiency of Esterase from Bacillus subtilis for Kinetic Resolution of l-Menthol through Semirational Design.

Enzymatic kinetic resolution is a promising way to produce l-menthol. However, the properties of the reported biocatalysts are still unsatisfactory and far from being ready for industrial application. Herein, a para-nitrobenzylesterase (pnbA) gene from Bacillus subtilis was cloned and expressed to produce l-menthol from d,l-menthyl acetate. The highest enantiomeric excess (ee) value of the product generated by pnbA was only approximately 80%, with a high conversion rate (47.8%) of d,l-menthyl acetate with the help of a cosolvent, indicating high catalytic activity but low enantioselectivity (E = 19.95). To enhance the enantioselectivity and catalytic efficiency of pnbA to d,l-menthyl acetate in an organic solvent-free system, site-directed mutagenesis was performed based on the results of molecular docking. The F314E/F315T mutant showed the best catalytic properties (E = 36.25) for d,l-menthyl acetate, with 92.11% ee and 30.58% conversion of d,l-menthyl acetate. To further improve the properties of pnbA, additional mutants were constructed based on the structure-guided triple-code saturation mutagenesis strategy. Finally, four mutants were screened for the best enantioselectivity (ee > 99%, E > 300) and catalytic efficiency at a high substrate concentration (200 g/L) without a cosolvent. This work provides several generally applicable biocatalysts for the industrial production of l-menthol.

The Influence of the Solid Solution Formation on Purification of L-Menthol from the Enantiomer Mixture by Three-Phase Crystallization.

Three-phase crystallization (TPC) was introduced in this study to purify L-menthol from menthol enantiomer mixtures in consideration of the formation of solid solutions. TPC is a new separation technology, which combines melt crystallization and vaporization to result in the desired crystalline product from a liquid mixture along with the unwanted components vaporized via the three-phase transformation by reducing temperature and pressure. The three-phase transformation conditions for the liquid menthol enantiomer mixtures were determined based on the thermodynamic calculations to direct the TPC experiments. A new model was proposed based on the mass and energy balances in consideration of the formation of the solid solutions to predict the yield and purity of the final L-menthol product during TPC. The yield and purity obtained from the TPC experiments were compared with those predicted by the model.

Composition and Anti-Helicobacter pylori Properties of Essential Oils Obtained from Selected Mentha Cultivars.

Helicobacter pylori infections are highly common amongst the global population. Such infections have been shown to be the cause of gastric ulcers and stomach carcinoma and, unfortunately, most cases are asymptomatic. Standard treatment requires antibiotics such as metronidazole or azithromycin to which many strains are now resistant. Mentha species have been used as a natural treatment for gastrointestinal diseases throughout history and essential oils (EOs) derived from these plants show promising results as potential antimicrobial agents. In this study, EOs obtained from the leaves and flowers of five cultivars of Mentha × piperita and M. spicata were examined by GC-MS. The investigated mints are representatives of four chemotypes: the menthol chemotype (M. × piperita 'Multimentha' and M. × piperita 'Swiss'), the piperitenone oxide chemotype (M. × piperita 'Almira'), the linalool chemotype (M. × piperita 'Granada'), and the carvone chemotype (M. spicata 'Moroccan'). The chemical composition of EOs from mint flowers and leaves was comparable with the exception of the Swiss cultivar. Menthol was the most abundant component in the leaves while menthone was highest in flowers. The H. pylori ATCC 43504 reference strain and 10 other H. pylori clinical strains were examined for their sensitivity to the EOs in addition to their major monoterpenoid components (menthol, menthone, carvone, dihydrocarvone, linalool, 1,8-cineole, and limonene). All tested mint EOs showed inhibitory activity against both the reference H. pylori ATCC 43504 strain (MIC 15.6-31.3 mg/L) and clinical H. pylori strains (MIC50/90 31.3-250 mg/L/62.5-500 mg/L). Among the reference monoterpenes, menthol (MIC50/90 7.8/31.3 mg/L) and carvone (MIC50/90 31.3/62.5 mg/L) had the highest anti-H. pylori activity, which also correlated with a higher activity of EOs containing these compounds (M. × piperita 'Swiss' and M. spicata 'Moroccan'). A synergistic and additive interaction between the most active EOs/compounds and antibiotics possibly points to a new plant-based anti-H. pylori treatment.

Cold shock precooling improves the firmness of chili pepper during postharvest storage and the molecular mechanisms related to pectin.

This research was conducted to explore the influence of cold shock on the firmness, a quality marker in chili pepper during 0-21 d storage and determine mechanism by cold shock impacted pectin. Chili peppers were exposed to cold shock precooling (0 ± 2 °C water/ice mixture) for 0-, 30-, 90- and 150-min, respectively. Results showed that cold shock alleviated loss of firmness throughout storage. Firmness was positively associated with sodium carbonate-soluble pectin content (r = 0.44), methylation degree of CDTA-soluble pectin (r = 0.82) and water-soluble pectin (WSP, r = 0.87), but negatively associated with WSP content (r = -0.76), and the activities of ß-galactosidase (r = -0.72) and pectinlyase (r = -0.74). Cold shock for 90 min was determined to be optimal. This study confirms the applicability of cold shock precooling to maintain firmness and thereby to extend the shelf life of chili pepper.

Catalytic asymmetric synthesis of cannabinoids and menthol from neral.

The selective conversion of natural or synthetic neral to (1R,6S)-trans-isopiperitenol would enable and expedite sustainable routes to menthol1,2 and cannabinoids3-5. However, this reaction has been considered impossible because its product is more reactive to the required acid catalysts than its starting material, resulting in several side products6-9. We now show that an unsymmetric, strong and confined chiral acid, a highly fluorinated imino-imidodiphosphate, catalyses this process with excellent efficiency and selectivity. Expanding the method to other α,ß-unsaturated aldehydes could enable access to new cannabinoids and menthol derivatives not readily accessible previously. Mechanistic studies suggest that the confined catalyst accomplishes this reaction by binding the product in an unreactive conformation, thereby preventing its decomposition. We also show how (1R,6S)-trans-isopiperitenol can be readily converted to pharmaceutically useful cannabinoids and menthol, each in the shortest and most atom-economic routes so far.

Promoting Activity of Terpenes on Skin Permeation of Famotidine.

Famotidine (FMT) is a competitive histamine-2 (H2) receptor antagonist that inhibits gastric acid secretion for the treatment of Gastroesophageal reflux disease. To study the promoting effect and mechanism of terpenes, including l-menthol, borneol, and geraniol, as chemical enhancers, FMT was used as a model drug. Attenuated total reflectance-Fourier transform IR spectroscopy (ATR-FTIR) and differential scanning calorimetry (DSC) were used to explore the effects of terpenes on the skin. Hairless mouse skin was mounted on Franz-type diffusion cell, and skin permeation experiment of FMT hydrogel was carried out. The results suggested that the thermodynamic activity influenced the permeability of the drug, and the main mechanism of terpenes to enhance skin permeation of the drug was based on increasing the fluidity of the intercellular lipids. Moreover, it was revealed that l-menthol simultaneously relaxed the packing structure and lamellar structure, whereas geraniol had a great influence on the lamellar structure only. Collectively, all terpenes had a promoting effect on skin permeation of FMT, indicating their potential as chemical enhancers to change the microstructure of stratum corneum and improve the permeation of FMT through the skin, and it has great potential to be used in transdermal formulations of FMT.

Activation mechanism of the mouse cold-sensing TRPM8 channel by cooling agonist and PIP2.

The transient receptor potential melastatin 8 (TRPM8) channel is the primary molecular transducer responsible for the cool sensation elicited by menthol and cold in mammals. TRPM8 activation is controlled by cooling compounds together with the membrane lipid phosphatidylinositol 4,5-bisphosphate (PIP2). Our knowledge of cold sensation and the therapeutic potential of TRPM8 for neuroinflammatory diseases and pain will be enhanced by understanding the structural basis of cooling agonist- and PIP2-dependent TRPM8 activation. We present cryo-electron microscopy structures of mouse TRPM8 in closed, intermediate, and open states along the ligand- and PIP2-dependent gating pathway. Our results uncover two discrete agonist sites, state-dependent rearrangements in the gate positions, and a disordered-to-ordered transition of the gate-forming S6-elucidating the molecular basis of chemically induced cool sensation in mammals.

DES-Based Biocatalysis as a Green Alternative for the l-menthyl Ester Production Based on l-menthol Acylation.

The deep eutectic solvent (DES)-based biocatalysis of l-menthol acylation was designed for the production of fatty acid l-menthyl ester (FME) using fatty acid methyl ester (FAME). The biocatalytic reaction was assisted by a lipase enzyme in the DES reaction medium. ւՒ-menthol and fatty acids (e.g., CA-caprylic acid; OA-oleic acid; LiA-linoleic acid; and LnA-linolenic acid) were combined in the binary mixture of DES. In this way, the DES provided a nonpolar environment for requested homogeneity of a biocatalytic system with reduced impact on the environment. The screening of lipase enzyme demonstrated better performance of immobilized lipase compared with powdered lipase. The performance of the biocatalytic system was evaluated for different DES compositions (type and concentration of the acid component). l-menthol:CA = 73:27 molar ratio allowed it to reach a maximum conversion of 95% methyl lauric ester (MLE) using a NV (Candida antarctica lipase B immobilized on acrylic resin) lipase biocatalyst. The recyclability of biocatalysts under optimum conditions of the system was also evaluated (more than 80% recovered biocatalytic activity was achieved for the tested biocatalysts after five reaction cycles). DES mixtures were characterized based on differential scanning calorimetry (DSC) and refractive index analysis.

Detoxification of monoterpenes by a family of plant glycosyltransferases.

Plant monoterpenes are challenging compounds, since they often act as solvents, and thus have both phytotoxic and antimicrobial properties. In this study an approach is developed to identify and characterize enzymes that can detoxify monoterpenoids, and thus would protect both plants and microbial production systems from these compounds. Plants respond to the presence of monoterpenes by expressing glycosyltransferases (UGTs), which conjugate the monoterpenoids into glycosides. By identifying these enzymes in a transcriptomics approach using Mentha × piperita, a family of UGTs was identified which is active on cyclic monoterpenoids such as menthol, and on acyclic monoterpenoids such as geranic acid. Other members of this family, from tomato, were also shown to be active on these monoterpenoids. In vitro and in vivo activity of different UGTs were tested with different substrates. We found that some glycosyltransferases significantly affect the toxicity of selected monoterpenoids in Escherichia coli, suggesting that glycosyltransferases can protect cells from monoterpenoid toxicity.

Improvement of solubility, stability and antioxidant activity of carotenoids using deep eutectic solvent-based microemulsions.

Natural carotenoids have been widely used as colorants and antioxidants in process of food, medicine, and cosmetic. However, the carotenoids have low bioactivity in vivo due to poor water-solubility. To enhance the solubility, stability and antioxidant activity of carotenoids, novel microemulsions (MEs) composed with deep eutectic solvents (DESs), tween 80 and water were developed as alternatives to organic solvents. The phase diagrams and physicochemical properties (viscosity, pH, and diameter) of the DES-based MEs were investigated at different temperatures. Then the solubility distribution, storage stability and DPPH free radical-scavenging activity of three carotenoids (astaxanthin, astaxanthin ester and lutein) in the MEs were evaluated. Compared with ethanol, methanol, and acetone, all the DES-based MEs studied significantly enhanced the solubility of the carotenoids due to the stronger hydrogen bonding and Van der Waals interactions. The highest solubilities of 0.27, 473.63, and 12.50 mg/mL for astaxanthin, astaxanthin ester and lutein, respectively, were observed in the MEs containing DES (DL-menthol:acetic acid = 1:2) at 35 â„ƒ. Moreover, astaxanthin ester can be well preserved in the MEs containing DES (DL-menthol:octanoic acid = 1:2) with a half-life of more than 69 days. In addition, the DPPH scavenging capacities of the three carotenoids in all the MEs were higher than the organic solvents. The results revealed that the DES-based MEs with low viscosity (<0.2 Pa•s) and mild acidic pH (4-5) are potential solvents for natural carotenoids in food processing and storage, medicine making, as well as biomaterials processing.

Purification of menthone and menthol from Mentha haplocalyx by suspension particle assisted solvent sublation, neuroprotective effect in vitro and molecular docking of menthol on amyloid-ß.

Suspension particle assisted solvent sublation was designed for the first time. The volatile monoterpenes in Mentha haplocalyx Briq were extracted using this method from a solution containing plant solid particles as the lower phase of solvent sublation. Under the optimum conditions of the solvent sublation (n-hexane/plant solid particles 20% ethanol-water solution system, pH 4, flotation time 30 min and air flow rate 30 mL/min), the extraction yields were 2.0 × 102 mg/kg, 9.5 × 101 mg/kg and 1.2 × 103 mg/kg for menthone, isomenthone and menthol, respectively. Compared with the traditional methods, the established suspension particle assisted solvent sublation might be an economical and efficient extraction method in some aspects. Through a cellular antioxidant activity experiment, menthol could alleviate H2O2-induced oxidative stress. Molecular docking was applied to simulate the molecular recognition process between amyloid-ß and menthol. The affinity energy of menthol was -12.59 kJ/mol, indicating that menthol might have neuroprotective activity and the potential to be an amyloid-ß inhibitor.

Selective terpene based therapeutic deep eutectic systems against colorectal cancer.

Cancer remains a major health problem worldwide, with colorectal cancer (CRC) being the third most incident and the second most lethal. Inflammation, on the other hand, has been highly associated with cancer development and maintenance, therefore, the reduction of the inflammatory microenvironment represents a promising therapeutic strategy. Deep eutectic systems (DES) are based on the combination of different components which together, at a certain molar ratio, present a deep decrease in their melting point compared with the individual compounds. When an active pharmaceutical ingredient is part of a DES it is designated by therapeutic deep eutectic system (THEDES). New THEDES combining terpenes with anticancer properties, such as safranal, menthol and linalool, with nonsteroidal anti-inflammatory drugs (NSAIDs), like ibuprofen, ketoprofen and flurbiprofen were produced. To evaluate THEDES anti-CRC therapeutic potential, their physico-chemical properties, bioavailability and bioactivity, were explored. Our results show that safranal:ibuprofen (3:1), safranal:ibuprofen (4:1) and menthol:ibuprofen (3:1) present promising therapeutic activity towards CRC cells due to a selective cytotoxic action towards cancer cells. menthol:ibuprofen (3:1) anti-proliferative action seems to be related with cell membrane disruption, reduction of the inflammation through the reduction of reactive oxygen species (ROS) production, and induction of apoptosis via caspase-3. On the other hand, safranal:ibuprofen (3:1) and safafranal:ibuprofen (4:1) seem to prevent tumour expansion only through the induction of apoptosis via caspase-3. Besides, these systems present an increase in ibuprofen permeability, with menthol:ibuprofen (3:1) increasing also ibuprofen's solubility thus its overall bioavailability. Knowing that cancer is a huge problematic situation that requires alternative therapies with less side effects, improved efficacy, associated with less costs and environmentally friendly, a new opportunity emerges for DES to be part of the pharmaceutical industry.

Structure and dynamics of hydrophobic deep eutectic solvents composed from terpene-fatty acids investigated by molecular dynamics simulation.

Hydrophobic deep eutectic solvents (HDES) composed of Terpenes and Fatty acids have recently been the subject of great interest for removing pollutants from an aqueous environment. Despite the specific application of Hydrophobic DES and the important role of molecular dynamics simulation in predicting the properties of these compounds, not many studies have been done on their intermolecular interaction. In this work, we performed molecular dynamics simulations for the eutectic mixture based on monoterpene (Menthol, Thymol) and Fatty acids such as Caprylic Acid, Decanoic Acid, Lauric Acid, and Myristic acid. Binary mixtures of Terpene and Fatty acids were prepared at molar ratios 1:1, and their properties were investigated at 323 K. We have carried out 50 ns in the ensemble NPT to understand thermo-physical properties that are largely dependent on the interaction between molecules. Here, the structural properties of the binary mixtures were evaluated, and the possible explanations for their thermo-physical properties have been presented. The interaction between Terpenes and Fatty acids was studied by the structural properties such as the atom-atom radial distribution functions (RDF) and the Hydrogen bonding network between species and Spatial distribution functions (SDF). The structural properties studies revealed that the interaction between Terpenes and Fatty acids decreased the reduction of the accumulation of Terpene molecules around each other in the binary mixtures. Evidence has been acquired that the interaction between the menthol molecules was mostly affected by the Fatty acids molecules. Also, the transport properties of the binary mixtures were explored using the mean-square displacement (MSD) for the centers of mass of molecules, self-diffusion of species, and vector reorientation dynamics (VRD) of bonds. The simulation results indicated that intermolecular interactions play an important role in the dynamic properties of species, and maintains the low melting point of the mixture.

Structural and dynamic properties of eutectic mixtures based on menthol and fatty acids derived from coconut oil: a MD simulation study.

The structural and dynamical properties of the binary mixture of Menthol (MEN) and Fatty acids (FAs) were investigated using molecular dynamics simulations. To this end, the relationship between the structural and dynamical properties of the eutectic mixtures of MEN and FAs with different molar percentages of FAs are studied. Structural properties of the eutectic mixtures were characterized by calculating the combined distribution functions (CDFs), radial distribution functions (RDFs), angular distribution functions (ADFs), hydrogen bonding networks, and spatial distribution functions (SDF). Additionally, our Results indicated robust interactions between menthol and Caprylic acid molecules Finally, the transport properties of the mixtures were investigated using the mean square displacement (MSD) of the centers of mass of the species, self-diffusion coefficients and vector reorientation dynamics (VRD) of bonds. Overall, our simulation results indicated that intermolecular interactions have a significant effect on the dynamic properties of species.

Menthol-assisted homogenous liquid-liquid microextraction for HPLC/UV determination of favipiravir as an antiviral for COVID-19 in human plasma.

Favipiravir is a promising antiviral agent that has been recently approved for treatment of COVID-19 infection. In this study, a menthol-assisted homogenous liquid-liquid microextraction method has been developed for favipiravir determination in human plasma using HPLC/UV. The different factors that could affect the extraction efficiency were studied, including extractant type, extractant volume, menthol amount and vortex time. The optimum extraction efficiency was achieved using 300 µL of tetrahydrofuran, 30 mg of menthol and vortexing for 1 min before centrifuging the sample for 5 min at 3467g. Addition of menthol does not only induce phase separation, but also helps to form reverse micelles to facilitate extraction. The highly polar favipiravir molecules would be incorporated into the hydrophilic core of the formed reverse micelle to be extracted by the non-polar organic extractant. The method was validated according to the FDA bioanalytical method guidelines. The developed method was found linear in the concentration range of 0.1 to 100 µg/mL with a coefficient of determination of 0.9992. The method accuracy and precision were studied by calculating the recovery (%) and the relative standard deviation (%), respectively. The recovery (%) was in the range of 97.1-103.9%, while the RSD (%) values ranged between 2.03 and 8.15 %. The developed method was successfully applied in a bioequivalence study of Flupirava® 200 mg versus Avigan® 200 mg, after a single oral dose of favipiravir administered to healthy adult volunteers. The proposed method was simple, cheap, more eco-friendly and sufficiently sensitive for biomedical application.

An amino acid ester of menthol elicits defense responses in plants.

KEY MESSAGE: Valine menthyl ester (ment-Val) acts as a plant defense potentiator for several crop species including soybean. Terpenoids, including menthol, exhibit potent abilities as plant defense potentiators in agriculture and horticulture. In the current study, we developed new terpene derivatives that consisted of menthol and various amino acids and that were expected to act as powerful plant defense potentiators. We used 6 amino acids possessing low-reactive sidechains to synthesize an array of amino acid ester of menthol (ment-aa) compounds. Transcript levels of two defense genes (pathogenesis-related protein 1 [PR1] and trypsin inhibitor [TI]) were evaluated in leaves of soybean plants 24 h after application of aquatic solution of menthol or menthol-aa, and revealed that the valine menthyl ester (ment-Val) alone elevated the transcript level of defense genes, and it did so only at the low dose of 1 µM, not at higher or lower doses tested. Moreover, it appeared that histone acetylation was involved in this effect. Application of ment-Val enabled soybean plants to sustain the increased transcript levels in their leaves for up to 3 days. Moreover, when ment-Val was additionally applied at day 4, at which time the transcript level had declined to the basal level, the transcript level was re-elevated, indicating the possibility that ment-Val could be repeatedly used to sustain pest control. Ment-Val was found to be chemically stable and effective for defense of several crop species. Collectively, these data show that terpenoid conjugates are useful for pest control instead of or in addition to pesticides.

Development of a green miniaturized quick, easy, cheap, effective, rugged, and safe approach in tandem with temperature-assisted solidification of floating menthol droplet for analysis of multiclass pesticide residues in milk.

A new green miniaturized quick, easy, cheap, effective, rugged, and safe approach was developed and used for the extraction of multiclass 16 pesticides in milk before gas chromatography analysis. The miniaturization of method reduced the consumption of chemical reagents and samples. Magnetic three-dimensional graphene was used as sorbent in the clean-up step. Choline chloride:lactic acid (1:2) natural deep eutectic solvent was used as desorption solvent. Temperature-assisted solidification of floating menthol drop was executed for preconcentration of analytes. The method parameters including sorbent, desorption solvent, sorption and desorption times, menthol amount, pH, and ionic strength were optimized. The limit of quantification and linear range were 0.03-0.38 and 0.03-250 µg/kg, respectively. The accuracy was assessed by recovery evaluation at the spike levels of 50 and 100 µg/kg, in the range of 61-119%, with relative standard deviations within 2.1-18.2%. The method was applied to the analysis of pasteurized low and high-fat bovine milk, and various pesticide residues were detected in the concentrations range of 1.24-4.68 µg/kg. Finally, the greenness of the procedure was evaluated using the Analytical Eco-Scale. This work represents the first application of hybrid miniaturized extraction/preconcentration using a natural deep eutectic solvent and menthol to analyze pesticides.

Synthesis, Antifungal Activity, 3D-QSAR, and Molecular Docking Study of Novel Menthol-Derived 1,2,4-Triazole-thioether Compounds.

A series of novel menthol derivatives containing 1,2,4-triazole-thioether moiety were designed, synthesized, characterized structurally, and evaluated biologically to explore more potent natural product-based antifungal agents. The bioassay results revealed that at 50 µg/mL, some of the target compounds exhibited good inhibitory activity against the tested fungi, especially against Physalospora piricola. Compounds 5b (R = o-CH3 Ph), 5i (R = o-Cl Ph), 5v (R = m,p-OCH3 Ph) and 5x (R = α-furyl) had inhibition rates of 93.3%, 79.4%, and 79.4%, respectively, against P. piricola, much better than that of the positive control chlorothalonil. Compounds 5v (R = m,p-OCH3 Ph) and 5g (R = o-Cl Ph) held inhibition rates of 82.4% and 86.5% against Cercospora arachidicola and Gibberella zeae, respectively, much better than that of the commercial fungicide chlorothalonil. Compound 5b (R = o-CH3 Ph) displayed antifungal activity of 90.5% and 83.8%, respectively, against Colleterichum orbicalare and Fusarium oxysporum f. sp. cucumerinum. Compounds 5m (R = o-I Ph) had inhibition rates of 88.6%, 80.0%, and 88.0%, respectively, against F. oxysporum f. sp. cucumerinu, Bipolaris maydis and C. orbiculare. Furthermore, compound 5b (R = o-CH3 Ph) showed the best and broad-spectrum antifungal activity against all the tested fungi. To design more effective antifungal compounds against P. piricola, 3D-QSAR analysis was performed using the CoMFA method, and a reasonable 3D-QSAR model (r2 = 0.991, q2 = 0.514) was established. The simulative binding pattern of the target compounds with cytochrome P450 14α-sterol demethylase (CYP51) was investigated by molecular docking.